Cancer Immunology, Immunotherapy

, Volume 60, Issue 10, pp 1447–1460 | Cite as

Pilot trial of interleukin-2 and zoledronic acid to augment γδ T cells as treatment for patients with refractory renal cell carcinoma

  • Joshua M. Lang
  • Mahazarin R. Kaikobad
  • Marianne Wallace
  • Mary Jane Staab
  • Dorothea L. Horvath
  • George Wilding
  • Glenn Liu
  • Jens C. Eickhoff
  • Douglas G. McNeel
  • Miroslav Malkovsky
Original article


Prior to the advent of VEGF-targeted therapies, renal cell carcinoma (RCC) was among the few solid tumors shown to respond to cytokine-based therapies such as interleukin-2 (IL-2) and interferon alpha. Previous work has shown that aminobisphosphonates, including zoledronic acid (ZA), are capable of activating human Vγ9 Vδ2 T cells in vitro, and these cells can be further expanded with IL-2. Moreover, these Vγ9 Vδ2 T cells have cytolytic activity in vitro to multiple human tumor cell lines. In the current report, we have conducted a pilot trial in patients with metastatic RCC, evaluating different doses of ZA in combination with low-dose IL-2 to determine whether combining these agents can promote in vivo proliferation of Vγ9 Vδ2 T cells and elicit an antitumor response. In 12 patients evaluated, no objective clinical responses were observed by RECIST criteria; however, two patients experienced prolonged stable disease. A modest increase in Vγ9 Vδ2 T-cell frequency could be detected by Day 8 of therapy in four of the nine patients who received at least one cycle of therapy, but not to the magnitude anticipated from preclinical models. Repeated administration of IL-2 and ZA resulted in both a diminished in vivo percentage of Vγ9 Vδ2 T cells as well as impaired expansion in vitro after the first cycle of therapy. These results suggest that repeated administration of IL-2 and ZA, at the doses and schedules used in this trial, may actually inhibit the proliferative capacity of Vγ9 Vδ2 T cell in patients with metastatic RCC.


Vγ9 Vδ2 lymphocyte Interleukin-2 Zoledronic acid Renal cell carcinoma 



We would like to thank Beth A. Fredricks for her invaluable technical assistance. This work was supported by an investigator-initiated award from Novartis Pharmaceuticals and by the University of Wisconsin Carbone Cancer Center Core Grant P30 CA014520. Dr. Lang is supported by NIH grant T32 CA009614.


  1. 1.
    Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics, 2010. CA Cancer J Clin 60(5):277–300PubMedCrossRefGoogle Scholar
  2. 2.
    Linehan WM, Zbar B, Bates BE, Zelefsky MJ, Yang JC (2001) Cancer of the kidney and ureter. In: DeVita VTHS, Rosenberg SA (eds) Cancer principles and practice of oncology, 6th edn. Lippincott Williams & Wilkins, Philadelphia, pp 1362–1396Google Scholar
  3. 3.
    Cohen HT, McGovern FJ (2005) Renal-cell carcinoma. N Engl J Med 353(23):2477–2490PubMedCrossRefGoogle Scholar
  4. 4.
    Sternberg CN, Davis ID, Mardiak J, Szczylik C, Lee E, Wagstaff J, Barrios CH, Salman P, Gladkov OA, Kavina A, Zarba JJ, Chen M, McCann L, Pandite L, Roychowdhury DF, Hawkins RE (2010) Pazopanib in locally advanced or metastatic renal cell carcinoma: results of a randomized phase III trial. J Clin Oncol 28(6):1061–1068PubMedCrossRefGoogle Scholar
  5. 5.
    Motzer RJ, Hutson TE, Tomczak P, Michaelson MD, Bukowski RM, Oudard S, Negrier S, Szczylik C, Pili R, Bjarnason GA, Garcia-del-Muro X, Sosman JA, Solska E, Wilding G, Thompson JA, Kim ST, Chen I, Huang X, Figlin RA (2009) Overall survival and updated results for sunitinib compared with interferon alfa in patients with metastatic renal cell carcinoma. J Clin Oncol 27(22):3584–3590PubMedCrossRefGoogle Scholar
  6. 6.
    Hudes G, Carducci M, Tomczak P, Dutcher J, Figlin R, Kapoor A, Staroslawska E, Sosman J, McDermott D, Bodrogi I, Kovacevic Z, Lesovoy V, Schmidt-Wolf IG, Barbarash O, Gokmen E, O’Toole T, Lustgarten S, Moore L, Motzer RJ (2007) Temsirolimus, interferon alfa, or both for advanced renal-cell carcinoma. N Engl J Med 356(22):2271–2281PubMedCrossRefGoogle Scholar
  7. 7.
    Escudier B, Bellmunt J, Negrier S, Bajetta E, Melichar B, Bracarda S, Ravaud A, Golding S, Jethwa S, Sneller V (2010) Phase III Trial of bevacizumab plus interferon alfa-2a in patients with metastatic renal cell carcinoma (AVOREN): final analysis of overall survival. J Clin Oncol 28(13):2144–2150PubMedCrossRefGoogle Scholar
  8. 8.
    Escudier B, Eisen T, Stadler WM, Szczylik C, Oudard S, Siebels M, Negrier S, Chevreau C, Solska E, Desai AA, Rolland F, Demkow T, Hutson TE, Gore M, Freeman S, Schwartz B, Shan M, Simantov R, Bukowski RM (2007) Sorafenib in advanced clear-cell renal-cell carcinoma. N Engl J Med 356(2):125–134PubMedCrossRefGoogle Scholar
  9. 9.
    Motzer RJ, Escudier B, Oudard S, Hutson TE, Porta C, Bracarda S, Grünwald V, Thompson JA, Figlin RA, Hollaender N, Urbanowitz G, Berg WJ, Kay A, Lebwohl D, Ravaud A (2008) Efficacy of everolimus in advanced renal cell carcinoma: a double-blind, randomised, placebo-controlled phase III trial. Lancet 372(9637):449–456PubMedCrossRefGoogle Scholar
  10. 10.
    Lang JM, Harrison MR (2010) Pazopanib for the treatment of patients with advanced renal cell carcinoma. Clin Med Insights Oncol 4:95–105PubMedGoogle Scholar
  11. 11.
    Motzer RJ, Mazumdar M, Bacik J, Russo P, Berg WJ, Metz EM (2000) Effect of cytokine therapy on survival for patients with advanced renal cell carcinoma. J Clin Oncol 18(9):1928–1935PubMedGoogle Scholar
  12. 12.
    Malaguarnera M, Ferlito L, Gulizia G, Di Fazio I, Pistone G (2001) Use of interleukin-2 in advanced renal carcinoma: meta-analysis and review of the literature. Eur J Clin Pharmacol 57(4):267–273PubMedCrossRefGoogle Scholar
  13. 13.
    RS Fisher RI, Fyfe G (2000) Long-term survival update for high-dose recombinant interleukin-2 in patients with renal cell carcinoma. Cancer J Sci Am 6(Suppl 1):S55–S57Google Scholar
  14. 14.
    Amato RJ, Morgan M, Rawat A (2006) Phase I/II study of thalidomide in combination with interleukin-2 in patients with metastatic renal cell carcinoma. Cancer 106(7):1498–1506PubMedCrossRefGoogle Scholar
  15. 15.
    Amato RJ, Malya R, Rawat A (2008) Phase II study of combination thalidomide/interleukin-2 therapy plus granulocyte macrophage-colony stimulating factor in patients with metastatic renal cell carcinoma. Am J Clin Oncol 31(3):237–243PubMedCrossRefGoogle Scholar
  16. 16.
    Lissoni P, Bordin V, Vaghi M, Fumagalli L, Bordoni A, Mengo S, Bucovec R, Fumagalli E, Malugani F, Ardizzoia A, Giani L, Gardani GS, Tancini G (2002) Ten-year survival results in metastatic renal cell cancer patients treated with monoimmunotherapy with subcutaneous low-dose interleukin-2. Anticancer Res 22(2B):1061–1064PubMedGoogle Scholar
  17. 17.
    Lanier LL, Ruitenberg J, Bolhuis RL, Borst J, Phillips JH, Testi R (1988) Structural and serological heterogeneity of gamma/delta T cell antigen receptor expression in thymus and peripheral blood. Eur J Immunol 18(12):1985–1992PubMedCrossRefGoogle Scholar
  18. 18.
    Wallace M, Malkovsky M, Carding SR (1995) Gamma/delta T lymphocytes in viral infections. J Leukoc Biol 58(3):277–283PubMedGoogle Scholar
  19. 19.
    Malkovsky M, Fisch P, Mackenzie D, Bartz SR, Radtke BE, Wallace M, Manning J, Colizzi V, Pauza CD (1992) Specificity and function of gamma delta T lymphocytes. Folia Biol (Praha) 38(5):293–306Google Scholar
  20. 20.
    Kato Y, Tanaka Y, Miyagawa F, Yamashita S, Minato N (2001) Targeting of tumor cells for human gammadelta T cells by nonpeptide antigens. J Immunol 167(9):5092–5098PubMedGoogle Scholar
  21. 21.
    Groh V, Rhinehart R, Secrist H, Bauer S, Grabstein KH, Spies T (1999) Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. Proc Natl Acad Sci U S A 96(12):6879–6884PubMedCrossRefGoogle Scholar
  22. 22.
    Bauer S, Groh V, Wu J, Steinle A, Phillips JH, Lanier LL, Spies T (1999) Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285(5428):727–729PubMedCrossRefGoogle Scholar
  23. 23.
    Viey E, Fromont G, Escudier B, Morel Y, Da Rocha S, Chouaib S, Caignard A (2005) Phosphostim-activated gamma delta T cells kill autologous metastatic renal cell carcinoma. J Immunol 174(3):1338–1347PubMedGoogle Scholar
  24. 24.
    Olive C, Nicol D, Falk MC (1997) Characterisation of gamma delta T cells in renal cell carcinoma patients by polymerase chain reaction analysis of T cell receptor transcripts. Cancer Immunol Immunother 44(1):27–34PubMedCrossRefGoogle Scholar
  25. 25.
    Kobayashi H, Tanaka Y, Yagi J, Toma H, Uchiyama T (2001) Gamma/delta T cells provide innate immunity against renal cell carcinoma. Cancer Immunol Immunother 50(3):115–124PubMedCrossRefGoogle Scholar
  26. 26.
    Kowalczyk D, Skorupski W, Kwias Z, Nowak J (1996) Activated gamma/delta T lymphocytes infiltrating renal cell carcinoma. Immunol Lett 53(1):15–18PubMedCrossRefGoogle Scholar
  27. 27.
    Kunzmann V, Bauer E, Feurle J, Weissinger F, Tony HP, Wilhelm M (2000) Stimulation of gammadelta T cells by aminobisphosphonates and induction of antiplasma cell activity in multiple myeloma. Blood 96(2):384–392PubMedGoogle Scholar
  28. 28.
    Sicard H, Ingoure S, Luciani B, Serraz C, Fournie JJ, Bonneville M, Tiollier J, Romagne F (2005) In vivo immunomanipulation of V gamma 9 V delta 2 T cells with a synthetic phosphoantigen in a preclinical nonhuman primate model. J Immunol 175(8):5471–5480PubMedGoogle Scholar
  29. 29.
    Casetti R, Perretta G, Taglioni A, Mattei M, Colizzi V, Dieli F, D’Offizi G, Malkovsky M, Poccia F (2005) Drug-induced expansion and differentiation of V gamma 9 V delta 2 T cells in vivo: the role of exogenous IL-2. J Immunol 175(3):1593–1598PubMedGoogle Scholar
  30. 30.
    Dieli F, Gebbia N, Poccia F, Caccamo N, Montesano C, Fulfaro F, Arcara C, Valerio MR, Meraviglia S, Di Sano C, Sireci G, Salerno A (2003) Induction of gammadelta T-lymphocyte effector functions by bisphosphonate zoledronic acid in cancer patients in vivo. Blood 102(6):2310–2311PubMedCrossRefGoogle Scholar
  31. 31.
    Dieli F, Vermijlen D, Fulfaro F, Caccamo N, Meraviglia S, Cicero G, Roberts A, Buccheri S, D’Asaro M, Gebbia N, Salerno A, Eberl M, Hayday AC (2007) Targeting human gamma delta T cells with zoledronate and interleukin-2 for immunotherapy of hormone-refractory prostate cancer. Cancer Res 67(15):7450–7457PubMedCrossRefGoogle Scholar
  32. 32.
    Meraviglia S, Eberl M, Vermijlen D, Todaro M, Buccheri S, Cicero G, La Mendola C, Guggino G, D’Asaro M, Orlando V, Scarpa F, Roberts A, Caccamo N, Stassi G, Dieli F, Hayday AC (2010) In vivo manipulation of Vgamma9Vdelta2 T cells with zoledronate and low-dose interleukin-2 for immunotherapy of advanced breast cancer patients. Clin Exp Immunol 161(2):290–297PubMedGoogle Scholar
  33. 33.
    Abe Y, Muto M, Nieda M, Nakagawa Y, Nicol A, Kaneko T, Goto S, Yokokawa K, Suzuki K (2009) Clinical and immunological evaluation of zoledronate-activated Vgamma9gammadelta T-cell-based immunotherapy for patients with multiple myeloma. Exp Hematol 37(8):956–968PubMedCrossRefGoogle Scholar
  34. 34.
    Sato K, Kimura S, Segawa H, Yokota A, Matsumoto S, Kuroda J, Nogawa M, Yuasa T, Kiyono Y, Wada H, Maekawa T (2005) Cytotoxic effects of gammadelta T cells expanded ex vivo by a third generation bisphosphonate for cancer immunotherapy. Int J Cancer 116(1):94–99PubMedCrossRefGoogle Scholar
  35. 35.
    Mariani S, Muraro M, Pantaleoni F, Fiore F, Nuschak B, Peola S, Foglietta M, Palumbo A, Coscia M, Castella B, Bruno B, Bertieri R, Boano L, Boccadoro M, Massaia M (2005) Effector gammadelta T cells and tumor cells as immune targets of zoledronic acid in multiple myeloma. Leukemia 19(4):664–670PubMedGoogle Scholar
  36. 36.
    Tourani JM, Lucas V, Mayeur D, Dufour B, DiPalma M, Boaziz C, Grise P, Varette C, Pavlovitch JM, Pujade-Lauraine E, Larregain D, Ecstein E, Untereiner M, Vuillemin E, Merran S, Andrieu JM (1996) Subcutaneous recombinant interleukin-2 (rIL-2) in out-patients with metastatic renal cell carcinoma. Results of a multicenter SCAPP1 trial. Ann Oncol 7(5):525–528PubMedGoogle Scholar
  37. 37.
    Wilhelm M, Kunzmann V, Eckstein S, Reimer P, Weissinger F, Ruediger T, Tony HP (2003) Gammadelta T cells for immune therapy of patients with lymphoid malignancies. Blood 102(1):200–206PubMedCrossRefGoogle Scholar
  38. 38.
    Zoledronic Acid [Package Insert]. Novartis pharmaceuticals. East Hanover, NJ (2001)Google Scholar
  39. 39.
    Caccamo N, Meraviglia S, Ferlazzo V, Angelini D, Borsellino G, Poccia F, Battistini L, Dieli F, Salerno A (2005) Differential requirements for antigen or homeostatic cytokines for proliferation and differentiation of human Vgamma9Vdelta2 naive, memory and effector T cell subsets. Eur J Immunol 35(6):1764–1772PubMedCrossRefGoogle Scholar
  40. 40.
    Kobayashi H, Tanaka Y, Yagi J, Osaka Y, Nakazawa H, Uchiyama T, Minato N, Toma H (2007) Safety profile and anti-tumor effects of adoptive immunotherapy using gamma-delta T cells against advanced renal cell carcinoma: a pilot study. Cancer Immunol Immunother 56(4):469–476PubMedCrossRefGoogle Scholar
  41. 41.
    Kobayashi H, Tanaka Y, Shimmura H, Minato N, Tanabe K (2010) Complete remission of lung metastasis following adoptive immunotherapy using activated autologous gammadelta T-cells in a patient with renal cell carcinoma. Anticancer Res 30(2):575–579PubMedGoogle Scholar
  42. 42.
    Bennouna J, Levy V, Sicard H, Senellart H, Audrain M, Hiret S, Rolland F, Bruzzoni-Giovanelli H, Rimbert M, Galea C, Tiollier J, Calvo F (2010) Phase I study of bromohydrin pyrophosphate (BrHPP, IPH 1101), a Vgamma9Vdelta2 T lymphocyte agonist in patients with solid tumors. Cancer Immunol Immunother 59(10):1521–1530PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Joshua M. Lang
    • 1
  • Mahazarin R. Kaikobad
    • 2
  • Marianne Wallace
    • 2
  • Mary Jane Staab
    • 3
  • Dorothea L. Horvath
    • 3
  • George Wilding
    • 1
  • Glenn Liu
    • 1
  • Jens C. Eickhoff
    • 4
  • Douglas G. McNeel
    • 1
    • 5
  • Miroslav Malkovsky
    • 2
  1. 1.Department of Medicine, Division of Hematology/OncologyUniversity of WisconsinMadisonUSA
  2. 2.Department of Medical Microbiology and ImmunologyUniversity of WisconsinMadisonUSA
  3. 3.Carbone Cancer CenterUniversity of WisconsinMadisonUSA
  4. 4.Department of Biostatistics and Medical InformaticsUniversity of WisconsinMadisonUSA
  5. 5.7007 Wisconsin Institutes for Medical ResearchMadisonUSA

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